Comparative translating ribosome affinity purification-RNAseq (TRAPseq) by lung cell-specific drivers

Translating ribosome affinity purification coupled to next-generation RNA sequencing permits profiling of specific cellular compartments. Here, we used well-characterized Cre and CreER drivers to lineage trace different cell populations in the developing lung with a ribosomal L10-GFP fusion protein. Two different anti-GFP antibodies were used to pull down ribosomes. mRNA was isolated, reversed transcribed, and processed using next-generation sequencing. Profiling showed distinct clustering between alveolar cell drivers and epithelial cell drivers, which were clearly distinguishable from raw mRNA input.